FlowCellect™ Multi-STAT Activation Profiling Kit

FlowCellect™ Multi-STAT Activation Profiling Kit

FlowCellect

25 Tests / Kit

Millipore’s FlowCellect™ Multi-STAT Activation Kit is designed to simultaneously detect the phosphorylation of the most commonly studied STAT proteins through cytokine or growth factor mediated pathway activation. The kit will also allow a researcher to quickly profile the status of constitutive activation of STAT1, STAT3, and STAT5 within a population of cells or primary tumors.
The ability to detect the activation of multiple STAT proteins simultaneously allows researchers to determine and profile the STAT signaling pathways within a cell type of interest. Alterations in these signaling profiles can be used to monitor the effect of a particular upstream treatment targeting the STAT pathways.
Antibodies against phospho-STAT1, 3, and 5A/B are provided in the kit to monitor the activation state of each STAT within a given cell type. U937 cells were stimulated with Gamma Interferon, IL-6, and GM-CSF simultaneously, to activate STAT1, STAT3, and STAT5A/B, respectively. When performing a three color stain, triple positive cell populations were detected.
Sufficient reagents are provided for 25 3-color samples. The kit includes all optimized fluorescently labeled antibodies and buffers necessary for cell preparation and analysis. Detailed assay instructions are included to assist in analysis and to ensure the correct cell concentration is obtained during acquisition of sample data.

STATs are transcription factors that can be phosphorylated normally by a cell surface receptor such as cytokine receptors, followed by dimerization and nuclear translocation where they bind to DNA and activate transcription of their target genes. STATs are involved in embryogenesis, cellular immunity, and cell proliferation [S Akira (1999)].

Many types of cancers both solid tumor and some blood based malignancies have been found to exhibit constitutive STAT activation [Ralf Buettner (2002)]. Interestingly, the activation within these cancer cells is generally a combination of STATs, typically STAT1, STAT3, and STAT5. This has led to an increase in research revolving around the complex family of STATs.

Oncogenesis and tumour progression are supported by alterations in cell signaling. Using flow cytometry, it is now possible to track and analyze signaling events in individual cancer cells [Jonathan M. Irish (2006)]. Single cell resolution of signaling molecules allows for a greater understanding of the elaborate processes that occur when cells transform and begin replicating uncontrollably.

Millipore’s FlowCellect STAT Activation kit includes three directly conjugated phospho specific antibodies: pSTAT1(Tyr701)-PerCP, pSTAT3(Tyr705)-Alexa 488, and pSTAT5A/B(Tyr694/Tyr699)-PE along with optimized fixation, permeabilization, wash and flow buffers to provide researchers with a complete solution for simultaneously detecting the activation of three of the most studied STAT proteins in their preferred cellular system.

Flow Cytometry

Human

• Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
• Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

Lot to lot in-process testing on all kit components

• 1. 20X pSTAT1 (Y701)-PerCP Monoclonal Antibody: (Part No. CS203319) 1 vial containing 150 µL antibody.
• 2. 20X pSTAT3 (Y705)-Alexa 488 Monoclonal Antibody: (Part No. CS203316) 1 vial containing 150 µL antibody
• 3. 20X pSTAT5A/5B (Y694/Y699)-PE Monoclonal Antibody: (Part No. CS203313) 1 vial containing 150 µL antibody.
• 4. 10X Wash Buffer: (Part No. CS202123) 1 bottle containing 13 mL buffer
• 5. Fixation Buffer: (Part No. CS202122) 1 bottle containing 13 mL buffer.
• 6. 5X Assay Buffer: (Part No. CS202124) 1 bottle containing 55 mL buffer
• 7. 1X Permeabilization Buffer: (Part No. CS203284) 2 bottles, each containing 15 mL.

Mouse

4-8°C for antibodies and buffers

• Fluorescent
• ¹²⁵I

FlowCellect™ Multi-STAT Activation Profiling Kit

Flow Cellect

ExpressPro

Monoclonal Antibody

Research